Zheng Yueliang, Li Gaoxiang, Shi Aili, Guo Junping, Xu Yingge, Cai Wenwei
Emergency and Critical Care Center, Department of Emergency Medicine, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, Zhejiang, China.
Rainbowfish Rehabilitation & Nursing School, Hangzhou Vocational & Technical College, Hangzhou, Zhejiang, China.
Heliyon. 2024 Oct 12;10(20):e39338. doi: 10.1016/j.heliyon.2024.e39338. eCollection 2024 Oct 30.
Acute lung injury (ALI) is a type of diffuse lung injury that seriously affects the survival of critically ill patients. MicroRNAs (miRNAs) can serve as promising therapeutic targets or offer insights for the development of potential therapeutic strategies against ALI. In our previous study, we demonstrated the protective effect of allicin in ALI, but the role of miRNAs in the alleviation of ALI by allicin remains unclear. This study aimed to investigate whether miRNAs mediate the effects of allicin on ALI. Cell viability and proliferation were determined using CCK-8 and EdU assays, respectively, while cellular apoptosis was analyzed by flow cytometry. The claudin-4 protein was detected by quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) and western blotting. The binding of miR-455 with claudin-4 was determined by bioinformatics analysis and validated by dual luciferase reporter assays. The lung wet/dry ratio of lipopolysaccharide (LPS)-treated rats was determined by hematoxylin and eosin (HE) and TUNEL staining of the pulmonary tissues. The levels of myeloperoxidase (MPO), interleukin (IL)-2, IL-6, and tumor necrosis factor (TNF)-α were determined by enzyme-linked immunosorbent assay (ELISA). We observed that allicin alleviated LPS-induced injury in A549 cells, and claudin-4 knockdown reversed the protective effect of allicin in ALI. Claudin-4 is a direct target of miR-455-3p, and miR-455-3p overexpression partially reversed the protective effect of allicin in LPS-treated A549 cells. Subsequent experiments confirmed that allicin protects against LPS-induced ALI by regulating the miR-455-3p/claudin-4 axis. The study revealed that the protective effect of allicin in ALI is mediated via miR-455-3p, which suppresses the expression of claudin-4.
急性肺损伤（ALI）是一种严重影响危重症患者生存的弥漫性肺损伤。微小RNA（miRNA）有望成为治疗靶点，或为开发针对ALI的潜在治疗策略提供思路。在我们之前的研究中，我们证明了大蒜素对ALI具有保护作用，但miRNA在大蒜素减轻ALI中的作用仍不清楚。本研究旨在探讨miRNA是否介导大蒜素对ALI的影响。分别使用CCK-8和EdU检测法测定细胞活力和增殖，同时通过流式细胞术分析细胞凋亡。通过定量实时逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测claudin-4蛋白。通过生物信息学分析确定miR-455与claudin-4的结合，并通过双荧光素酶报告基因检测法进行验证。通过苏木精-伊红（HE）染色和肺组织的TUNEL染色测定脂多糖（LPS）处理大鼠的肺湿/干比。通过酶联免疫吸附测定（ELISA）测定髓过氧化物酶（MPO）、白细胞介素（IL）-2、IL-6和肿瘤坏死因子（TNF）-α的水平。我们观察到大蒜素减轻了LPS诱导的A549细胞损伤，而claudin-4基因敲低逆转了大蒜素对ALI的保护作用。claudin-4是miR-455-3p的直接靶点，miR-455-3p过表达部分逆转了大蒜素对LPS处理的A549细胞的保护作用。随后的实验证实，大蒜素通过调节miR-455-3p/claudin-4轴来预防LPS诱导的ALI。该研究表明，大蒜素对ALI的保护作用是通过miR-455-3p介导的，miR-455-3p可抑制claudin-4表达。
